Probing enantioselective substrate specificity for macrocyclization by 6-deoxyerythronolide thioesterase
LE3 .A278 2015
Bachelor of Science
The synthesis, isolation, and purification of enzymatic substrates were achieved by an eleven-step enantioselective synthesis, which included the Crimmins asymmetric aldol reaction and its corresponding acetate aldol reaction. All substrates and intermediates were characterized by 1H and 13C nuclear magnetic resonance (NMR) and high resolution mass spectrometry (HRMS). Biochemical assays are to be performed in vitro to characterize the ability of the 6-deoxyerythronolide B (6-dEB) recombinant purified thioesterase (TE) to macrocyclize the substrates, which will be analyzed via liquid chromatography-mass spectrometry (LCMS) to determine the specificity of, and probe the factors that contribute to, macrocyclization. These experiments are predicted to further our understanding regarding the effects of methyl substituents on the macrocyclization using DEBS TE to generate novel analogues that may be used for further testing and drug development.
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